RESUMO
Mitogen-activated protein kinase kinase 6 (MKK6) and activator protein-1 (AP-1) are two of the essential regulatory proteins in the p38 mitogen-activated protein kinase (MAPK) pathway, which participates in the innate immune response to bacterial infections. In this study, molluscan MKK6 (AwMKK6) and AP-1 (AwAP-1) genes were cloned and identified from Anodonta woodiana. The open reading frame (ORF) of AwMKK6 encodes for a putative polypeptide sequence of 345 amino acids containing a conserved serine/threonine protein kinase (S_TKc) domain, a SVAKT motif and a DVD domain. AwAP-1 consists of 294 amino acids including a typical nuclear localization signal (NLS), a Jun domain and a basic region leucine zipper (BRLZ) domain. Quantitative real-time PCR analysis showed that both AwMKK6 and AwAP-1 were widely expressed in all selected tissues of A. woodiana and their transcript levels in hemocytes were significantly upregulated when challenged with Aeromonas hydrophila and lipopolysaccharide (LPS). Additionally, the signaling molecules of the AwMKK6/AwAP-1 pathway including AwTLR4, AwMyD88, AwTRAF6, AwMEKK1, AwMEKK4, AwASK1, AwTAK1 and Awp38 mRNA expression showed a stronger responsiveness to LPS challenge in hemocytes of A. woodiana. RNA interference (RNAi) experiments indicated that the silencing of AwMKK6 or AwAP-1 could decrease the mRNA expression levels of immune effectors (AwTNF, AwLYZ and AwDefense). Subcellular localization studies suggested that AwMKK6 and AwAP-1 were distributed throughout the cells and nucleus, respectively, and their overexpression could significantly enhance the transcriptional activities of AP-1-Luc in HEK293T cells. These findings suggest that MKK6 and AP-1 play a major role in the host defense response to bacterial injection, which may make contributions to a better understanding of the immune function of the p38 MAPK pathway in mollusks.
Assuntos
Anodonta , Aminoácidos , Animais , Anodonta/genética , Células HEK293 , Humanos , Imunidade Inata/genética , Lipopolissacarídeos/farmacologia , RNA Mensageiro/metabolismo , Fator de Transcrição AP-1/genéticaRESUMO
Transferrin, a member of the iron binding superfamily protein, plays an extremely important role in the transport of iron in the biological process of cells. The result of preliminary proteomic study on E. sinensis hemocytes infected Spiroplasma eriocheiris showed the expression of transferrin (EsTF) and ferrin (EsFe) significantly changed. In addition, other reports have confirmed that transferrin, ferritin and iron are involved in the immune response of hosts. In order to validate the immune function of EsTF, the whole length of EsTF was successfully amplified by the gene cloning and RACE technique. The results showed that the full-length cDNA of the EsTF gene was 2748 bp, including a 2193 bp open reading frame which encodes 730 amino acids. The result of bioinformatics analysis showed EsTF contains two highly conserved TR_FER domains. Evolutionary analysis showed that EsTF has a close genetic relationship with other TFs of invertebrates. In addition, EsTF mRNA was highly transcripted in nerve and intestine tissues, followed by hemocytes. The expression of EsTF, EsFe1 and EsFe2 increased after exogenous supplemental of iron under the concentration of 100â¯nmol/L in water. After exogenous supplement of iron and injection with S. eriocheiris, these three gene transcription of mRNA levels were higher than that of PBS group, while lower than the S. eriocheiris group and the iron group. Besides, the copy number of S. eriocheiris in the experimental group was significantly reduced, and the death rate decreased. As can be seen, iron made transferrin and ferritin return to normal levels during the infection of S. eriocheiris and help the host maintain normal immunity levels to resist S. eriocheiris. These results further demonstrated that EsTF, EsFe1, EsFe2 and iron play a role in the immune defense mechanism of the crabs to resist S. eriocheiris infection.
Assuntos
Braquiúros/genética , Braquiúros/imunologia , Imunidade Inata/genética , Ferro/metabolismo , Spiroplasma/fisiologia , Transferrina/genética , Transferrina/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Hemócitos/imunologia , Hemócitos/microbiologia , Filogenia , Proteômica , Distribuição Aleatória , Transferrina/químicaRESUMO
Cyprinid herpesvirus 2 (CyHV-2) is a pathogen of herpesviral hematopoietic necrosis disease of crucian carp. Our study aimed to investigate the molecular mechanisms and immune response at the mRNA and protein levels in head kidney during CyHV-2 infection. Three days after infection with CyHV-2, 7085 differentially expressed genes were identified by transcriptome sequencing, of which 3090 were up-regulated and 3995 were down-regulated. And 338 differentially expressed proteins including 277 up-regulated and 61 down-regulated were identified using tandem mass tag labeling followed by liquid chromatography tandem mass spectrometry. Notably, 128 differentially co-expressed genes at mRNA and protein levels (cDEGs) were reliably quantified, including 86 co-up-regulated and 42 co-down-regulated. In addition, 10 cDGEs in the above pathways were selected for qRT-PCR to confirm the validity of the transcriptome and proteome changes by showing that RIG-I, MDA5, LGP2, FAS, PKR and PKZ up-regulated and Integrin α, Integrin ß2, NCF2 and NCF4 down-regulated. This indicated that after CyHV-2 infection, the herpes simplex infection pathway, RIG-I like receptor signaling pathway, necroptosis pathway and p53 signaling pathway were activated and the phagosome pathway was suppressed. Our findings reveal the pathogenesis and the host immune mechanism of CyHV-2 infection of crucian carp.
Assuntos
Carpas , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteoma , Transcriptoma , Animais , Proteínas de Peixes/metabolismo , Perfilação da Expressão Gênica , Herpesviridae/fisiologia , Infecções por Herpesviridae/imunologia , Proteômica , Distribuição AleatóriaRESUMO
A ras-related nuclear protein (Ran) protein was obtained from Macrobrachium rosenbergii, named MrRan. Phylogenetic analysis results showed that MrRan was clustered in one group together with other crustaceans. Tissue distribution analysis revealed that MrRan was expressed mainly in gill, intestine and stomach, and expressed weakly in muscle. The MrRan expression levels in gill and hemocyte of prawns were significantly up-regulated after challenged by Spiroplasma eriocheiris. The copy number of S. eriocheiris in MrRan dsRNA injection group was significantly less than control groups during infection. Meanwhile, silencing MrRan obviously increased the survival rate of prawns. The subcellular localization experiment suggested that recombinant MrRan was mainly located in the nucleus, and relatively weak in the cytoplasm. Finally, over-expression in Drosophila S2 cell indicated that MrRan could increase copies of S. eriocheiris and decrease of cell viability. The present study suggested that MrRan participated in regulating the phagocytosis of S. eriocheiris in M. rosenbergii.
Assuntos
Imunidade Inata/genética , Palaemonidae/genética , Palaemonidae/imunologia , Proteína ran de Ligação ao GTP/genética , Proteína ran de Ligação ao GTP/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Brânquias/imunologia , Hemócitos/imunologia , Filogenia , Spiroplasma/fisiologia , Proteína ran de Ligação ao GTP/químicaRESUMO
Interfacial thermal resistance is the main barrier restricting the heat dissipation of thermal management materials in electronic equipment. The interface structure formed by covalent bonding is an effective way to promote interfacial heat transfer. Herein, an integrated composite with multi-aspect covalent bonding beneficial for heat transmission is constructed by polyimide (PI) polymerization with maleimide modified graphene nanosheets (M@GNS). The interfacial structure with low thermal resistance built by covalent bonding and oriented graphene arrangement initiated by the coating process makes the in-plane thermal conductivity of the composite as high as 16.10 W m-1 K-1. Finite element simulation and 1000 bending tests are carried out to further verify the performance advantages of the integrated structure in the internal thermal diffusion and long-term use of the composite. M@GNS/PI with integrated structure provides extra heat transfer channels for heat dissipation, possibly providing an effective way to address the traditional thermal accumulation issue of electronic devices.
RESUMO
Heat dissipation efficiency and electromagnetic interference (EMI) shielding performance are vital to integration, miniaturization, and application of electronic devices. Flexible and designable polymer-based composites are promising candidates but suffer from unavoidable interfacial thermal resistances, anisotropic thermal conductivity, and low shielding of EMI limiting application. Herein, multifunctional epoxy resin (EP)-based composites with an interconnected carbon fibers (CFs) network structure containing a low thermal resistance interfacial were prepared by high-temperature calcination and infiltration. The coherent heat and electron transfer pathways constructed with self-oriented CFs cloth connected by carbon nanotubes (CNTs) converted from leaf-shaped zeolitic imidazolate frameworks (ZIF-L) and stable magnetic property provided by cobalt nanoparticles contained in the CNTs made composites to an integrated in-plane thermal conductivity of up to 7.50 W m-1 K-1, a through-plane thermal conductivity of 1.96 W m-1 K-1, and an EMI shielding effectiveness of 38.4 dB. Furthermore, the mechanical properties of CFs and the junction effect of CNTs endowed the composites with stability of mechanical property, thermal conductivity, and EMI shielding effectiveness after multiple bendings. The finite element simulation further verified the advantage of CFs network linked by CNTs on heat transfer. This work provides the desired design for the construction of a multifunctional polymer-based composite used in advanced electronic equipment.
RESUMO
The effect of hydronephrosis, a common complication of metastatic colorectal cancer (CRC), on the treatment outcome and prognosis of locally advanced or metastatic CRC remains to be elucidated. The present study investigated the clinical characteristics, outcomes, and prognoses of patients with locally advanced or metastatic colorectal cancer (CRC) with hydronephrosis. Clinical data of patients with locally advanced or metastatic CRC who were attending Peking University Shenzhen Hospital and Shenzhen Cancer Hospital between January 2016 and December 2020 were retrospectively collected. A total of 52 patients with hydronephrosis based on CT or MRI findings were selected, and their clinical characteristics, treatment outcomes, and survival times were analyzed. Of the 52 patients, 33 were male (63.5%), and the median age was 49 years (range, 31-76 years). A total of 15 (28.8%) patients with CRC had synchronous hydronephrosis and the remaining 37 patients had metachronous hydronephrosis. Ureters were either compressed by peritoneal or abdominal cavity metastatic lymph nodes in 34 cases (65.4%) or by direct tumor invasion in 18 cases (34.6%). However, objective response rate (ORR) was higher in the group in which ureters were compressed by peritoneal or abdominal cavity metastatic lymph nodes; ORR, disease control rate and median progression-free survival (PFS) between the two groups were not statistically different. (P>0.05). The median survival period was only 27.0 months (95% CI, 20.549-33.451) in patients complicated with malignant hydronephrosis. Univariate and multivariate analyses showed that CA19-9 might be a prognostic factor for locally advanced and metastatic CRC patients with hydronephrosis. Metachronous metastatic CRC has a high incidence rate of complicated hydronephrosis. Targeted drugs in combination with chemotherapy improve the treatment efficacy and prognosis of patients. Notably, the present study found that CA19-9 level might be a prognostic factor in CRC patients with hydronephrosis.